Formalin fixed paraffin embedded tumor tissue from mice treated for 2 days were sliced manually with a manual microtome (Leica, RM2155) to 5 μm depth. H&E staining was preformed using Gill's II Hematoxylin (Sigma-Aldrich, GHS216) and Eosin Y (VWR, 95057-848). For IHC, slides were stained with cleaved caspase 3 antibody (Cell Signaling Technology, 9661), and counterstained with Gill's II Hematoxylin. Slides were visualized using a Leica DMi8 automated microscope and also scanned by HistoWiz for low-power visualization.
Gill s 2 hematoxylin
Manufactured by Merck Group
Sourced in United States
Gill's II Hematoxylin is a laboratory staining reagent used in histological and cytological procedures. It serves as a nuclear stain, highlighting the nuclei of cells in prepared tissue samples or cell preparations. The product provides a consistent and reliable staining outcome, enabling clear visualization and identification of cellular structures under microscopic examination.
Automatically generated - may contain errors
2 protocols using gill s 2 hematoxylin
1
Histological Analysis of Tumor Tissue
2
Immunohistochemical Analysis of NeuN in Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For immunohistochemistry (IHC) the formalin-fixed tissue was processed and embedded in paraffin blocks. Coronal sections were cut at 5 μm thickness and placed on adhesive glass slides.
The sections were air dried and then baked for 20 minutes at 60˚C. Sections were then deparaffinized and hydrated using a series of xylenes and alcohol changes. Antigen retrieval was performed using an Ethylenediaminetetraacetic acid (EDTA) buffer in a steamer followed by rinsing with phosphate buffered saline (PBS).
Primary antibody was applied, and signal amplification was performed using a horseradish peroxidase-labelled detection kit (NeuN antibody #MAB377, MilliporeSigma, USA). Following the application of each solution, it was ensured that the samples were thoroughly rinsed with the use of PBS.
Finally, the Diaminobenzidine (DAB, Sigma, USA) reaction was performed allowing visualization of the primary antibodies and subsequent analysis using optical microscopy. Slides were rinsed with distilled water and counterstained with Gill's II Hematoxylin (Sigma, USA) to make the pia and superficial layers more visible and allow for pia thickness measurements. Slides were dehydrated and cover slipped for permanent mounting.
The sections were air dried and then baked for 20 minutes at 60˚C. Sections were then deparaffinized and hydrated using a series of xylenes and alcohol changes. Antigen retrieval was performed using an Ethylenediaminetetraacetic acid (EDTA) buffer in a steamer followed by rinsing with phosphate buffered saline (PBS).
Primary antibody was applied, and signal amplification was performed using a horseradish peroxidase-labelled detection kit (NeuN antibody #MAB377, MilliporeSigma, USA). Following the application of each solution, it was ensured that the samples were thoroughly rinsed with the use of PBS.
Finally, the Diaminobenzidine (DAB, Sigma, USA) reaction was performed allowing visualization of the primary antibodies and subsequent analysis using optical microscopy. Slides were rinsed with distilled water and counterstained with Gill's II Hematoxylin (Sigma, USA) to make the pia and superficial layers more visible and allow for pia thickness measurements. Slides were dehydrated and cover slipped for permanent mounting.
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